Methods for identifying treatments that reduce the actions of substances of abuse and addiction

Abstract

Based on the discovery that MBLAC1 is a specific, high-affinity target for Ceftriaxone (Cef), MBLAC1 may be used for identifying treatments for addiction to substances of abuse. Methods for identifying therapeutic agents for treatment of addiction to a substance of abuse include using an assay to determine if a test agent is capable of binding to MBLAC1 or disrupting binding between MBLAC1 protein and Cef, and identifying such a test agent as a candidate therapeutic agent for treatment of addiction to a substance of abuse. MBLAC knock-out (KO) animals, methods of use thereof, and kits are used for identifying a therapeutic agent that reduces the actions of at least one substance of abuse. Methods also include using cellular extracts from tissue or cultured cells taken from wild-type (WT) MBLAC1 and MBLAC1 KO animals for screening for novel, Cef-like molecules in vitro, and using cells from a MBLAC1 KO animal to test for Cef-like actions of a test molecule.

What is claimed is: 
     
       1. A method for identifying therapeutic agents for treatment of addiction to a substance of abuse that require MBLAC1 to reduce actions of the substance of abuse, the method comprising the steps of:
 (a) providing at least one test agent, MBLAC1 protein or MBLAC1-expressing cells, and Ceftriaxone (Cef); and (b) using an assay to determine whether the at least one test agent is capable of disrupting binding between MBLAC1 protein and Cef, wherein a test agent capable of disrupting binding between MBLAC1 protein and Cef is identified as a candidate therapeutic agent for treatment of addiction to a substance of abuse; and ((c) administering the candidate therapeutic agent and the substance of abuse to at least one MBLAC1 knock-out (KO) animal and to at least one wild-type (WT) MBLAC1 animal and subsequently subjecting the animals to at least one test selected from the group consisting of: a locomotor assay, a withdrawal assay, a sensitization assay, consisting of: a locomotor assay, a self-administration assay, a reinstatement to drug assay, an analysis of white matter changes, and an analysis of changes in GLTI expression, 
 wherein if the candidate therapeutic agent reduces or eliminates the actions of the substance of abuse in the WT MBLAC1 animal but not in the MBLAC1 KO animal, the candidate therapeutic agent is a therapeutic agent that requires the presence of MBLAC1 to reduce actions of a substance of abuse. 
 
     
     
       2. The method of  claim 1 , wherein the test agent is capable of disrupting binding between MBLAC1 protein and Cef and/or binds to MBLAC1 protein with an affinity of K D =2 μM or less. 
     
     
       3. The method of  claim 1 , wherein the substance of abuse is selected from the group consisting of: cocaine, amphetamine, morphine, ethanol, methamphetamine, clorazepate, cathinones, bath salts, heroin, nicotine, alcohol, ketamine, and MDMA. 
     
     
       4. The method of  claim 1 , wherein the MBLAC1 protein is human MBLAC1 protein. 
     
     
       5. The method of  claim 1 , wherein the at least one test agent is a β-lactam antibiotic. 
     
     
       6. The method of  claim 1 , wherein a library of test agents comprises the at least one test agent. 
     
     
       7. The method of  claim 6 , wherein the library consists essentially of β-lactam structures. 
     
     
       8. The method of  claim 6 , wherein the library is an organic molecule library or a peptide library. 
     
     
       9. The method of  claim 1 , wherein the assay of step (b) is selected from the group consisting of: microcalorimetry, surface plasmon resonance, backscattering interferometry, radioligand binding assay, and an assay that can detect binding of unlabeled small molecules and proteins. 
     
     
       10. The method of  claim 1 , wherein the animals are rodents.